• Title/Summary/Keyword: isozyme profiles

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A New Strategy for Determining Optimum pH of Isozymes

  • Yoon, Kil-Joong
    • Bulletin of the Korean Chemical Society
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    • v.25 no.7
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    • pp.997-1002
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    • 2004
  • A hydrogenperoxide sensor containing peroxidase extracted from horseradish was constructed and pH effect on its sensing ability was investigated. Current profiles of the biosensor with pH and the electrophoretic analysis showed that horseradish peroxidase consists of two isozymes. Assuming that it is a hypothetical twoisozyme mixture, the current profiles were deconvoluted into two Gaussians. Application of the new Michaelis-Menten equation connoting pH concept to this system enabled to find all the related dissociation constants of the isozyme-substrates and the isozyme-proton complexes and to determine pHs for the maximal isozyme activities.

The allopolyploid origin of Euphorbia stevenii and E. boöphthona (Euphorbiaceae)

  • Ki-Ryong PARK
    • Korean Journal of Plant Taxonomy
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    • v.52 no.4
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    • pp.219-225
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    • 2022
  • To elucidate the ancestry of the allopolyploids E. stevenii and E. boöphthona, I examined eleven isozyme loci and 24 morphological characters from 28 populations representing five related Euphorbia species from Australia. According to an analysis of genetic and morphological data, three diploid species differentiated recently, but two independent polyploid species are estimated to have differentiated a relatively long time ago. Fixed heterozygosity for most isozymes in E. stevenii and E. boöphthona strongly suggests that these two species are allopolyploids rather than autopolyploids. The isozyme profiles of E. stevenii indicate that it is an allopolyploid that evolved from interspecific hybridization between the diploid E. tannensis and unidentified or extinct tetraploid species. In addition, isozyme patterns strongly suggest that E. stevenii was one of the ancestors of E. boöphthona. However, E. boöphthona showed a large number of fixed alleles that were not detected in any other Australian Eremophyton species. The most likely hypothesis for the origin of E. boöphthona is that it was formed by hybridization and chromosomal doubling between an extinct diploid species and the hexaploid E. stevenii.

Studies on Protein Profiles and Isozymes in Germinating Seeds (종자발아에 있어서 Protein Profile과 Isozyme에 관한 연구)

  • 권오용
    • Journal of Plant Biology
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    • v.17 no.4
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    • pp.143-156
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    • 1974
  • The purpose of this experiment was to study one side of germination physiology based on that protein profiles and protease relating to protein metabolism, that peroxidase, catalase, $\alpha$-amylase, $\beta$-amylase, and malate dehydrogenase involved in the carbohydrate metabolism of seed germination. All these experiments were divided into the two groups with and without acetone treatment, and were carried out. The protein bands of each germinating stage between the groups treated with and without acetone showed certain basic pattern in polyacrylamide gel disc electrophoresis. However, there was a little difference in the number of protein band, optical density, and migration velocity between two groups. The isozyme bands of peroxidase, and catalase between two groups in polyacrylamide gel disc electrophoresis did not show the numeral difference, but the optical density of certain germinating stage treated with acetone was higher than the group untreated with it and it showed their enzyme activity. The $\alpha$-amylase and $\beta$-amylase activities which involved in starch metabolism of seed germination were higher in the treated group than the other. On one hand, the protease activity of hydrolase occurred in the seeds for germination was also higher, more or less in the treated group than in the other. The isozyme band pattern of malate dehydrogenase in TCA cycle of energy metabolism pathway was very different between two groups growing for 72 hours with and without acetone treatment in cellulose acetate electrophoresis. It indicated that two isozyme bands of malate dehydrogenase was high. Consequently these experimental results mentioned above indicated that acetone treatment before sowing had an effect on dissolving certain complexed lipid substance involved in the seed coats, the activity of carbohydrate hydrolase increased with water absorption which was most comfortable in its germination, dissolved glycerin and fatty acid became certain energy source, and they stimulated the acceleration of respiration metabolism.

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Electrochemical Determination of the Optimum pH of HRP (전기화학적 방법에 의한 HRP의 최적 pH 도출)

  • Yoon, Kil-Joong
    • Analytical Science and Technology
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    • v.16 no.6
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    • pp.504-508
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    • 2003
  • A carbon paste electrode was constructed with peroxidase extracted from Horseradish and the variation of the response of the sensor with pH was investigated. Current profiles showed two highest sensitivities at two pH values respectively. In addition, two bands were observed in the electrophoretic expansion. A coincidence of the two experimental results added support to the possibility that the biosensor has two different isozymes. Assuming that current profiles are the sum of two gaussians, we deconvoluted them and determined the optimum pH of peroxidase isozymes.

Mechanisms of Chilling Tolerance in Relation to Antioxidative Enzymes in Rice

  • Kuk, Yong-In;Shin, Ji-San;Whang, Tay-Eak;Guh, Ja-Ock
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.5
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    • pp.341-351
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    • 2002
  • In order to examine the mechanistic basis for differential sensitivities to chilling and subsequent recovery between two rice (Oryza sativa L.) cutivars, a chilling-tolerant japonica type (Ilpumbyeo) and a chilling-susceptible indica type (Taebaekbyeo), changes of physiological responses and antioxidant enzymes were investigated. Both cultivars at 3 leaf stage were exposed at a low temperature of $5^{\circ}C$ for 3 days and subsequently recovered in a growth chamber at a $25^{\circ}C$ for 5 days with 250 mmol $m^{-2}$ $s^{-1}$. Physiological parameters such as leaf fresh weight, relative water content, cellular leakage, lipid peroxidation, and chlorophyll a fluorescence showed that the chilling tolerant cultivar had a high tolerance during chilling. However, the chilling-susceptible cultivar revealed severe chilling damages. The chilling-tolerant cultivar was also faster in recovery than the chilling-susceptible cultivar in all parameters examined. We analyzed the activity and isozyme profiles of four antioxidant enzymes which are: superoxide dismutase (SOD), caltalase (CAT), ascorbate peroxidase (APX), and glutation reductase (GR). We observed that chilling-tolerance was due to a result of the induced or higher antioxidant enzyme system, CAT and APX in leaves and SOD, CAT, APX, and GR in roots. Especially, we observed the most significant differences between the chilling-tolerant cultivar and -susceptible cultivar in CAT and APX activity. Also in isozyme profiles, CAT and APX band intensity in the chilling-tolerant cultivar was distinctively higher than in the chilling-susceptible cultivars during chilling and recovery. Thus, the cold stability of CAT and APX are expected to contribute to a tolerance mechanism of chilling in rice plants. In addition, the antioxidative enzymes activity in roots may be more important than in that of leaves to protect chilling damage on rice plants.

Somatic Hybrids by Electro-Protoplast Fusion between N. tabacum and N. glutinosa (담배(N. tabacum)와 N. glutinosa 종간 원형질체 융합식물의 생성)

  • 김준철;최성진
    • KSBB Journal
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    • v.5 no.2
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    • pp.175-182
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    • 1990
  • Protoplasts, isolated from leaf of N. tabacum NR-/SR+ and N. glottnosa were electrofused and divided with a plating efficiency of 30∼35% in AAPI 9M medium. Green callus lines were selected in protoplast-derived colonies on MSNO3 selection medium with 1.2mg/ml streptomycin sulfate on the basis of nitrate reductase proficiency and streptomycin resistance. Four putative hybrid plant lines regenerated from the green callus lines had intermediate morphology between that of parents with respect to floral shape, corolla length and ovate leaf blade. Zymograms of leaf peroxidase and esterase from these putative hybrid plant lines showed isozyme profiles derived from both parents and also, they exhibited additional and lost bands. Cytological analysis of two putative hybrid plant lines gave chromosome counts of 2n=66 in L22 and 2n=54 in L44 which were less than the expected number of N. tabacum(2n=48) and N. glutinosa(2n=24).

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Genetic Diversity Studies and Identification of Molecular and Biochemical Markers Associated with Fusarium Wilt Resistance in Cultivated Faba Bean (Vicia faba)

  • Mahmoud, Amer F.;Abd El-Fatah, Bahaa E.S.
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.11-28
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    • 2020
  • Faba bean (Vicia faba L.) is one of the most important legume crops in Egypt. However, production of faba bean is affected by several diseases including fungal diseases. Fusarium wilt incited by Fusarium oxysporum Schlecht. was shown to be the most common wilt disease of faba bean in Assiut Governorate. Evaluation of 16 faba bean genotypes for the resistance to Fusarium wilt was carried out under greenhouse conditions. Three molecular marker systems (inter-simple sequence repeat [ISSR], sequence related amplified polymorphism [SRAP], and simple sequence repeat [SSR]) and a biochemical marker (protein profiles) were used to study the genetic diversity and detect molecular and biochemical markers associated with Fusarium wilt resistance in the tested genotypes. The results showed that certain genotypes of faba bean were resistant to Fusarium wilt, while most of the genotypes were highly susceptible. The percentage of disease severity ranged from 32.83% in Assiut-215 to 64.17% in Misr-3. The genotypes Assiut-215, Roomy-3, Marut-2, and Giza2 were the most resistant, and the genotypes Misr-3, Misr-1, Assiut-143, Giza-40, and Roomy-80 performed as highly susceptible. The genotypes Assiut-215 and Roomy-3 were considered as promising sources of the resistance to Fusarium wilt. SRAP markers showed higher polymorphism (82.53%) compared with SSR (76.85%), ISSR markers (62.24%), and protein profile (31.82%). Specific molecular and biochemical markers associated with Fusarium wilt resistance were identified. The dendrogram based on combined data of molecular and biochemical markers grouped the 16 faba bean genotypes into three clusters. Cluster I included resistant genotypes, cluster II comprised all moderate genotypes and cluster III contained highly susceptible genotypes.

Activities and Isozyme Profiles of Antioxidant Enzymes in Intercellular Compartment of Overwintering Barley Leaves

  • Baek, So-Hyeon;Kwon, In-Sook;Park, Tae-Il;Yun, Song-Joong;Kim, Jin-Key;Choi, Kyeong-Gu
    • BMB Reports
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    • v.33 no.5
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    • pp.385-390
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    • 2000
  • Overwintering plants should survive the various biotic and abiotic stresses that occur during winter. Previous studies indicated that active oxygen species are involved in freezing, dehydration, anoxia and pathogen infections. As the importance of the events that occur in the intercellular compartment became apparent in disease resistance, we examined the nature of intercellular antioxidant enzymes in order to access their possible involvement in the winter hardiness of barley. The levels of intercellular peroxidase, catalase, and SOD activities on the unit protein basis were 394, 18, and 9% of those of cellular activities, respectively. Major intercellular peroxidase isoforms consisted of four neutrals and four basic forms; whereas major cellular isoforms were two basic forms. Out of the two major catalase isoforms a higher molecular weight form was predominantly abundant in both cellular and intercellular compartments. Among the five major cellular SOD isoforms, three were also present in the intercellular compartment. The presence of substantial amounts of intercellular antioxidant enzymes in overwintering barley leaves may suggest the involvement of these enzymes in the tolerance mechanism to the various stresses that occur during winter.

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Ripening Related Changes in Hot Pepper Fruit Cell Walls Structural Alterations of Cell Wall Polysaccharides and Separation of Galactosidase Isozymes (고추의 성숙에 따른 세포벽 다당류의 변화와 ${\beta}-Galactosidase$ Isozymes의 분리)

  • Kim, Soon-Dong;Kang, Meung-Su;Kim, Kwang-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.157-163
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    • 1985
  • Various cell wall polysaccharides and related enzyme activities in hot pepper fruit were determined at different stages of maturity. The uronic acid content of cell walls decreased between immature green and turning stage fruit and then increased by red ripe stage. In contrast, cellulose content of cell walls changed only a little during ripening. Total neutal sugar content of cell wall material decreased 50% and galactose content of the walls decreased about 80% by the turning stage. Polygalacturonase and ${\beta}-galactosidase$ activities, as well as total hemicellulose from isolated cell walls of ripening hot pepper fruit were studied using gel filtration chromatography. Polygalacturonase activity was not detectable but 5 isozymes of ${\beta}-galactosidase$ were resolved. The activities of the enzymes were relatively high and gel filtration showed that they differed in molecular weight. Hemicellulose content decreased during ripening and softening. The molecular weight profiles shifted from high molecular weight to low molecular weight polymers during ripening. The changes in cell walls that may be associated with fruit softening involve the alteration of hemicellulose prior to the degradation of wall-bound uronic acid. It is suggested that the decrease in cell wall galactose involved changes in turnover of new cell wall components.

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